Objective:To investigate the effects of matrix metalloproteinase 10(MMP10)on the proliferation,migration,invasion,and lymph node metastasis of oral squamous cell carcinoma(OSCC)cells,and to explore the underlying mechanisms.Methods:Gene expression profile data and survival data of head and neck squamous cell carcinoma(HNSCC)tissues and adjacent non-cancerous tissues were downloaded from The Cancer Genome Atlas(TCGA).Protein expression profile data of HNSCC tissues and adjacent non-cancerous tissues were downloaded from the Clinical Proteomic Tumor Analysis Consortium(CPTAC)database,and select the data of OSCC from HNSCC.The data from public database were used to analyze the differences in MMP10 expression at the mRNA and protein level between HNSCC/OSCC tissues and adjacent non-cancerous tissues,and to analyze the impact of MMP10 expression level on the survival rate of HNSCC patients,as well as the impact of human papillomavirus(HPV)on MMP10 gene expression in HNSCC patients.MMP10 expression level in 52 pairs of OSCC tissue samples and adjacent non-cancerous tissue samples(from the First Affiliated Hospital of Fujian Medical University)was measured by real-time quantitative PCR.After silencing MMP10 expression using siRNA(siMMP10-1291 and siMMP10-1336),the effect of MMP10 on the proliferation ability of OSCC cells was detected by CCK-8 assay and colony formation assay.The effects of MMP10 on the migration and invasion abilities of OSCC cells was assessed by wound-healing assay and transwell assay,respectively.To verify the effect of MMP10 knocking down on lymph node metastasis of OSCC cells in vivo,we constructed a nude mouse tongue cancer orthotopic model by shRNA(shMMP10-1291 and shMMP10-1336).Differential expression genes associated with MMP10 were screened from the gene expression profile data of OSCC patients in the TCGA database,and pathway enrichment analysis was performed using the GSEA software to further analyze the correlation between MMP10 and genes from the significantly enriched pathway.Results:Compared with adjacent non-cancerous tissues,the expression level of MMP10 in HNSCC and OSCC tissues was significantly increased(P<0.000 1),and HNSCC patients with high MMP10 expression had a lower survival rate(HR=1.32,P=0.04).HNSCC patients in the HPV-group had higher MMP10 gene expression levels(P<0.000 1),but their survival rate was significantly lower than that of HNSCC patients in the HPV+group(HR=0.48,P=0.004).Downregulating MMP10 expression could inhibit the migration,invasion and lymph node metastasis in vivo of OSCC cells.In OSCC patients,differentially expressed genes associated with MMP10 were significantly enriched in the Wnt signaling pathway(NES=1.505,P=0.029),and multiple genes in the Wnt signaling pathway,including WNT3,WNT3A,WNT5A,and WNT5B,were positively correlated with the expression level of MMP10.Conclusion:MMP10 promotes the invasion,metastasis and lymph node metastasis in vivo of OSCC cells by regulating the Wnt signaling pathway.

Objective:To investigate the effects of matrix metalloproteinase 10(MMP10)on the proliferation,migration,invasion,and lymph node metastasis of oral squamous cell carcinoma(OSCC)cells,and to explore the underlying mechanisms.Methods:Gene expression profile data and survival data of head and neck squamous cell carcinoma(HNSCC)tissues and adjacent non-cancerous tissues were downloaded from The Cancer Genome Atlas(TCGA).Protein expression profile data of HNSCC tissues and adjacent non-cancerous tissues were downloaded from the Clinical Proteomic Tumor Analysis Consortium(CPTAC)database,and select the data of OSCC from HNSCC.The data from public database were used to analyze the differences in MMP10 expression at the mRNA and protein level between HNSCC/OSCC tissues and adjacent non-cancerous tissues,and to analyze the impact of MMP10 expression level on the survival rate of HNSCC patients,as well as the impact of human papillomavirus(HPV)on MMP10 gene expression in HNSCC patients.MMP10 expression level in 52 pairs of OSCC tissue samples and adjacent non-cancerous tissue samples(from the First Affiliated Hospital of Fujian Medical University)was measured by real-time quantitative PCR.After silencing MMP10 expression using siRNA(siMMP10-1291 and siMMP10-1336),the effect of MMP10 on the proliferation ability of OSCC cells was detected by CCK-8 assay and colony formation assay.The effects of MMP10 on the migration and invasion abilities of OSCC cells was assessed by wound-healing assay and transwell assay,respectively.To verify the effect of MMP10 knocking down on lymph node metastasis of OSCC cells in vivo,we constructed a nude mouse tongue cancer orthotopic model by shRNA(shMMP10-1291 and shMMP10-1336).Differential expression genes associated with MMP10 were screened from the gene expression profile data of OSCC patients in the TCGA database,and pathway enrichment analysis was performed using the GSEA software to further analyze the correlation between MMP10 and genes from the significantly enriched pathway.Results:Compared with adjacent non-cancerous tissues,the expression level of MMP10 in HNSCC and OSCC tissues was significantly increased(P<0.000 1),and HNSCC patients with high MMP10 expression had a lower survival rate(HR=1.32,P=0.04).HNSCC patients in the HPV-group had higher MMP10 gene expression levels(P<0.000 1),but their survival rate was significantly lower than that of HNSCC patients in the HPV+group(HR=0.48,P=0.004).Downregulating MMP10 expression could inhibit the migration,invasion and lymph node metastasis in vivo of OSCC cells.In OSCC patients,differentially expressed genes associated with MMP10 were significantly enriched in the Wnt signaling pathway(NES=1.505,P=0.029),and multiple genes in the Wnt signaling pathway,including WNT3,WNT3A,WNT5A,and WNT5B,were positively correlated with the expression level of MMP10.Conclusion:MMP10 promotes the invasion,metastasis and lymph node metastasis in vivo of OSCC cells by regulating the Wnt signaling pathway.

Objective:To explore the different metabolites in the follicular fluids (FFs) of polycystic ovary syndrome (PCOS) patients and non-PCOS patients and their effects on the maturation of mouse oocytes and the developmental potential of in vitro fertilization (IVF) embryos. Methods:The clinical data were collected for the retrospective cohort study. Animal experiments were conducted in a randomized controlled trial. This study included PCOS ( n=71) and non-PCOS ( n=70) patients who underwent the first IVF or intracytoplasmic sperm injection (ICSI) cycle in Shanghai JIAI Genetics & IVF institute from June 2019 to June 2020. The patients' FFs were collected and the clinical data from these patients were analyzed. Raman spectroscopy analysis technology was used to detect differences in the metabolic spectra of FFs between the two groups. Mouse GV phase oocytes were placed in FFs from PCOS patients and non-PCOS patients for in vitro maturation (IVM) culture respectively, then the matured mouse oocytes were collected for IVF. The effects of differential metabolites in FFs on mouse oocyte maturation and embryonic development were further explored. The Raman spectrum was also applied to identify the differences of the IVM spent culture media. Results:The MⅡ rate [82.19% (886/1 078)] and day 3 available embryo rate [51.30% (553/1 078)] from PCOS group were significantly lower than those of the non-PCOS group [85.85% (625/728), P=0.038; 53.30% (388/728), P=0.042]. However, there were no significant differences between the two groups in the cumulative clinical pregnancy rate and the cumulative live birth rate (all P>0.05). Raman was capable of distinguishing PCOS from non-PCOS FFs. The characteristic Raman displacement difference between the two groups is mainly concentrated in the 600-1 000 cm -1, as well as 1 168 cm -1, 1 344 cm -1, 1 440 cm -1, 1 504 cm -1, 1 632 cm -1 and 1 664 cm -1. The Raman characteristic shift database showed that the different metabolites of the two sets of FFs samples were mainly concentrated in protein, lipids, free nucleic acis, glucose, cholesterol, carotenoids, and amino acids. Mouse oocyte IVM results showed that the PCOS-FF group had a lower MⅡ rate [49.04% (77/157)] than that of non-PCOS group [65.07% (95/146), P=0.005). IVF results showed the PCOS-FF group had a significantly lower cleavage rate [46.75% (36/77)] than that of non-PCOS group [63.16% (60/95), P=0.031], but there was no significant difference in the blastocyst rate between the two groups ( P>0.05). Conclusion:Differential metabolites detected by Raman spectrum in the PCOS FFs may cause defected maturation of the oocytes, leading to infertility, and Raman spectroscopy is an effective approach towards PCOS diagnosis and the identification of metabolomics differences.

Objective:To explore the different metabolites in the follicular fluids (FFs) of polycystic ovary syndrome (PCOS) patients and non-PCOS patients and their effects on the maturation of mouse oocytes and the developmental potential of in vitro fertilization (IVF) embryos. Methods:The clinical data were collected for the retrospective cohort study. Animal experiments were conducted in a randomized controlled trial. This study included PCOS ( n=71) and non-PCOS ( n=70) patients who underwent the first IVF or intracytoplasmic sperm injection (ICSI) cycle in Shanghai JIAI Genetics & IVF institute from June 2019 to June 2020. The patients' FFs were collected and the clinical data from these patients were analyzed. Raman spectroscopy analysis technology was used to detect differences in the metabolic spectra of FFs between the two groups. Mouse GV phase oocytes were placed in FFs from PCOS patients and non-PCOS patients for in vitro maturation (IVM) culture respectively, then the matured mouse oocytes were collected for IVF. The effects of differential metabolites in FFs on mouse oocyte maturation and embryonic development were further explored. The Raman spectrum was also applied to identify the differences of the IVM spent culture media. Results:The MⅡ rate [82.19% (886/1 078)] and day 3 available embryo rate [51.30% (553/1 078)] from PCOS group were significantly lower than those of the non-PCOS group [85.85% (625/728), P=0.038; 53.30% (388/728), P=0.042]. However, there were no significant differences between the two groups in the cumulative clinical pregnancy rate and the cumulative live birth rate (all P>0.05). Raman was capable of distinguishing PCOS from non-PCOS FFs. The characteristic Raman displacement difference between the two groups is mainly concentrated in the 600-1 000 cm -1, as well as 1 168 cm -1, 1 344 cm -1, 1 440 cm -1, 1 504 cm -1, 1 632 cm -1 and 1 664 cm -1. The Raman characteristic shift database showed that the different metabolites of the two sets of FFs samples were mainly concentrated in protein, lipids, free nucleic acis, glucose, cholesterol, carotenoids, and amino acids. Mouse oocyte IVM results showed that the PCOS-FF group had a lower MⅡ rate [49.04% (77/157)] than that of non-PCOS group [65.07% (95/146), P=0.005). IVF results showed the PCOS-FF group had a significantly lower cleavage rate [46.75% (36/77)] than that of non-PCOS group [63.16% (60/95), P=0.031], but there was no significant difference in the blastocyst rate between the two groups ( P>0.05). Conclusion:Differential metabolites detected by Raman spectrum in the PCOS FFs may cause defected maturation of the oocytes, leading to infertility, and Raman spectroscopy is an effective approach towards PCOS diagnosis and the identification of metabolomics differences.

Objective To investigate the changes of erythrocyte parameters and the value of differential diagnosis in pregnant women with β-mediterranean anemia.Methods A total of 300 pregnancy women from July 2014 to December 2015 in Center Hospital of Longgang were recruited in this study,100 pregnant women with β-mediterranean anemia in β-mediterranean anemia pregnancy group,100 healthy pregnant women in normal pregnancy group,100 pregnant women with iron deficiency anemia in iron deficiency anemia pregnancy group.Mean red cell volume(MCV),mean erythrocyte hemoglobin(MCH),reticulocyte percentage(Ret%) were detected and compared in the three groups.Results Compared with the normal pregnancy group and iron deficiency anemia pregnancy group,the MCV,MCH significantly reduced,and Ret% significantly rised in the β-mediterranean anemia pregnant group,the differences were significant(P<0.05).The best cut-off value of Ret% was 1.7% in differential diagnosis of β-mediterranean anemia pregnancy and iron deficiency anemia pregnancy,the sensitivity was 63.00%,the specificity was 74.00%,the area under of receiver operating characteristic curve was 0.841.The sensitivity of joint detection including MCV,MCH and Ret% in differential diagnosis of β-mediterranean anemia pregnancy and iron deficiency anemia pregnancy was 84.00%,the specificity was 90.00%.Conclusion MCV,MCH and Ret% in pregnancy women with β-mediterranean anemia changes significant compared with normal pregnancy group and iron deficiency anemia pregnancy group,the joint detection including MCV,MCH and Ret% could significantly improve the differential diagnosis of β-mediterranean anemia and iron deficiency anemia in pregnancy women.

Objective To determine the contents of four effective constituents in Euphorbiae Semen decoction;To provide evidence for Euphorbiae Semen decoction into clinical application. Methods Established quantitative analysis multi-components by single marker method was used to determine the contents of diterpenoids constituents, such as euphorbia storoid, euphorbia factor L2, and euphorbia factor L3. HPLC method was used to determine the contents of aesculetin. Results Contents of euphorbia storoid, euphorbia factor L2, and euphorbia factor L3 in smashed Euphorbiae Semen decoction were 0.015 9%, 0.005 9% and 0.024 1%, respectively. However, the contents of the above three constituents could not be detected in whole Euphorbiae Semen decoction. The content of aesculetin (0.693 6%) in smashed Euphorbiae Semen decoction was more than that in whole Euphorbiae Semen decoction (0.288 2%). Conclusion Decoction digestion effect of diterpenoids constituents in Euphorbiae Semen decoction is not good. Decocting with water is not suitable for the clinical application of Euphorbiae Semen for purgation and diuresis. Aesculetin in smashed Euphorbiae Semen decoction has good decoction digestion effect, in which clinical use for antisepsis and anti-inflammation is effective.

Objective To study the relationship between reactive oxygen species (ROS) as well as total antioxidant capacity ( TAC ) within follicle fluid and body mass index ( BMI ) in patients with polycystic ovary syndrome (PCOS).Methods All patients enrolled in this study were infertile women receiving in vitro fertilization-embryo transfer (IVF-ET) treatment.55 PCOS patients were divided into over-weight group ( n =23 ) and non-over-weight group ( n =32 ).Another 55 age-matched non-PCOS women were also divided into control group ( n =30) and overweight group ( n =25 ).Plasma sex hormone,triglycerides,and total cholesterol were determined.On oocyte retrieval day after ovarian stimulation,ROS and TAC in follicular fluid were assayed.Results Subjects in over-weight and PCOS over-weight groups had higher triglycerides than those in control and PCOS non-over-weight groups [ ( 1.9 ±1.1,1.7 ± 0.9,1.0 ± 0.5,1.2 ± 0.7 ) mmol/L,respectively,all P＜0.05],so did total cholesterol [ ( 4.8 ± 1.2,5.2 ± 1.1,4.0 ± 0.6) mmol/L,respectively,all P＜0.05].In PCOS over-weight group,ROS and ROS/TAC within follicular fluid were ( 35.4 ± 6.7 ) RLU/S and 39.8 ± 22.0,both were higher than those in the other 3 groups ( all P＜0.05).TAC [ (0.8 ± 0.5 ) Mm] was lower in PCOS over-weight group than that among the other 3 groups( all P＜0.05 ).ROS/TAC was higher in PCOS non-over-weight group than that of control group ( 26.5 ± 14.5 vs 14.2 ± 12.5,P＜0.05 ).Univariate analysis showed that both ROS and ROS/TAC within follicular fluid in PCOS patients were positively correlated with BMI ( r =0.34 and r =0.32,both P＜0.05 ).Conclusion Abnormal oxidative stress exists in follicular fluid of PCOS patients,and the oxidative stress parameters show positive correlation with BMI.